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MicroRNA-145 inhibits human papillary cancer TPC1 cell proliferation by targeting DUSP6.

International journal of clinical and experimental medicine (2015-08-27)
Yifan Gu, Dengfeng Li, Qifeng Luo, Chuankui Wei, Hongming Song, Kaiyao Hua, Jialu Song, Yi Luo, Xiaoyu Li, Lin Fang
ABSTRACT

MicroRNAs (miRNAs) are small, non-coding RNAs that modulate gene expression by negatively regulating the stability or translational efficiency of their target mRNAs. The aim of this study was to investigate the expression of microRNA-145 (miR-145) in human papillary thyroid cancer and its potential function. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to determine the expression level of miR-145 in ten papillary thyroid cancer and adjacent normal specimens. The function of miR-145 overexpression on the proliferation of human TPC1 thyroid cancer cells was conducted by MTT assays and by colony-formation assays. Western blot was used to validate the impact of miR-145 on the protein expression of the target gene. Luciferase reporter assays were employed to validate a putative target of miR-145. MiR-145 expression was relatively decreased in papillary thyroid cancer specimens compared with adjacent normal tissues (P<0.05). MTT assays and colony-formation assays showed that overexpression of miR-145 suppressed TPC1 cell growth. Luciferase assays using a reporter carrying a putative miR-145 target site in the 3' untranslated region of DUSP6 revealed that miR-145 directly targets DUSP6. Overexpression of miR-145 led to downregulation of DUSP6 at protein level as assessed by Western blot. Targeted knockdown of DUSP6 by siRNA significantly inhibited the proliferation of TPC1 cells. The overexpression of miR-145 inhibited TPC1 cellular growth by targeting DUSP6; this finding implies a better understanding of initiation and progression of papillary thyroid cancer.

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